Separation of the antimicrobial lipopeptide from acidic methanol extract by using reversed-phase HPLC. (A) Chromatogram profile of acidic methanol extract showed four peaks (F1-F4) and among these fractions F2 (lipopeptide) and F4 (bacteriocin-like peptide) showed antimicrobial activity. Diagonal red line indicates the gradient of solvent B; inset picture shows the re-chromatogram profile of lipopeptide fraction (F2). (B) Tricine–SDS-PAGE of the purified bacteriocin-like peptide. (C) Direct overlay of the SDS-PAGE gel demonstrating a clear inhibition zone against test strain S. aureus (indicated with arrow).